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Development of multiplex methylation-specific PCRfor detecting methylated/un-methylated PTEN promoterin breast cancer patients using a single closed tube |
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| รหัสดีโอไอ | |
| Creator | 1. Primariadewi Rustamadji 2. Azamris 3. Nunung Ainur Rahmah 4. Elsa Septiani Putri 5. Rosita Wati 6. Nadya Shifani 7. Yeremiah Rubin Camin 8. Endry Nugroho Prasetyo 9. Kartini Eriani 10. Elvan Wiyarta 11. Desriani |
| Title | Development of multiplex methylation-specific PCRfor detecting methylated/un-methylated PTEN promoterin breast cancer patients using a single closed tube |
| Publisher | Research and Development Office, Prince of Songkla University |
| Publication Year | 2565 |
| Journal Title | Songklanakarin Journal of Science an Technology (SJST) |
| Journal Vol. | 44 |
| Journal No. | 3 |
| Page no. | 869-875 |
| Keyword | PTEN, Methylation, recombinant plasmid, breast cancer, detection, diagnosis, multiple |
| URL Website | https://rdo.psu.ac.th/sjst/index.php |
| ISSN | 0125-3395 |
| Abstract | PTEN is a suppressor gene that methylation in its promoter is potentially used for diagnosis and prognosis for breastcancer and other cancer. We aim to develop a simple, fast, and economical Multiple Methylation-Specific PCR (MMSP) PTEN,recognizing a PTEN promoter gene for methylation and un-methylation developed in one single closed tube. This trueexperimental study uses a recombinant plasmid prepared with a TA cloning system. For the development of PTEN with MultipleMSPs, two formulations for MMSP PTEN were designed and compared, in which the best was to study specificity, limit ofdetection, reproducibility, and repeatability. The methods were validated with a clinical sample that identified its methylationstatus using Methylation-Specific PCR (MSP). As a result, the best formulation for MMSP PTEN was formulation 2, whichcomprises of 1:3 methylation and un-methylation primer, with only high specificity for detecting its target and high sensitivity of1.5625%, while its limit detection was 0.00032 ng/ยตl. It has high reproducibility and repeatability, which showed consistent datain the experiments with clinical samples in this activity. This simple molecular evidence aids in detecting biomarkers that arepotentially used for the diagnosis and prognosis of cancer and the epidemiological study of methylated PTEN promoters. |
